Method for Recombinant Expression and Purification of Vitellogenin in Channa sinensis

Abstract

The invention relates to a method for recombinant expression and purification of vitellogenin of S. sinensis, which relates to the technical field of expression and purification of fusion proteins. It provides a method based on the provitolin gene of S. sinensis. It clones part of the gene and recombines the plasmid in vitro, and transfers it into Escherichia coli for prokaryotic production. Expression, the IPTG induction temperature is 16°C to obtain high-efficiency fusion expression, and the expressed target protein exists in a soluble state, and the target protein accounts for more than 25% of the total bacterial soluble protein. The fusion protein MNP-VTG was obtained by applying purification technology, and the tagged MNP was separated from the fusion protein by sucrase digestion to obtain the target protein VTG monomer. This method is simple to express and purify VTG with high efficiency, because it is soluble, the separation is simpler than inclusion bodies, and no renaturation is required. The obtained recombinant VTG provides a stable and uniform raw material basis for subsequent applications. The recombinant protein can be used for biological activity and function research, and corresponding antibodies can also be prepared for detection research.

Description

technical field [0001] The invention relates to the technical field of fusion protein expression and purification methods, in particular to a method for expressing and purifying vitellogenin fusion proteins of the Chinese snakehead snakehead. Background technique [0002] Vitellogenin VTG is the precursor of vitellin in the egg development of oviparous animals. It is expressed by liver cells under the induction of estrogen, and the protein produced provides the hemolymph system to enter the ovary and enter the egg to mature into vitellin. Provides nutrients for egg development. As a reproductive nutritional protein, VTG is generally induced by hormones during yolk development (1. Bownes M. The roles of juvenile hormone, ecdysone and the ovary in the control of Drosophila vitellogenesis [J]. JInsect Physiol, 1989, 35: 409 -413; 2.C.M.Campbell DRI.Hormonal control of vitellogenesis inhypophysectomized winter flounder(Pseudopleuronectes americanusWalbaum)[J].Gen Comp Endocrino...

AI Technical Summary

Problems solved by technology

Moreover, all the reported recombinant VTGs currently exist in inclusion bodies, and renaturation is required to obtain active recombinant proteins, and the renaturation rate cannot reach 100%, and the separation and purification procedures are increased.

The presence of recombinant protein in soluble form is more advantageous than inclusion body form, and there is no report on the expression of VTG recombinant expression in soluble form.

[0007] There is no report about the recombinant expression and purification method of vitellogenin in Chinese snakehead snakehead

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